Reactivity of genotypically distinct hepatitis B virus surface antigens in 10 commercial diagnostic kits available in Japan

Hepatitis B virus (HBV) surface antigen (HBsAg) is one of the most important serological markers of current HBV infection. However, there are significant antigenic variations of HBsAg caused by genotypic diversity as well as mutation of the HBV genome. It is predictable that amino acid substitutions occurring in the HBsAg "a" determinant of a particular HBV genotype will affect the sensitivity of some diagnostic kits, since all the diagnostic kits currently available utilize monoclonal and/or polyclonal antibodies against the "a" determinant. One possible concern is that there may be a significant variation in the sensitivity of HBsAg diagnostic kits to HBsAg encoded by HBV of different genotypes, which might result in a failure to detect HBsAg of a particular HBV genotype. In this study, we assessed the reactivity of HBsAg specimens derived from three different HBV genotypes (A, B, and C) that are prevalent in Japan by 10 commercially available EIA (enzyme immunoassay), CLIA (chemiluminescent immunoassay), and CLEIA (chemiluminescent enzyme immunoassay) diagnostic kits. Specimens included both clinical samples and recombinant HBsAg. Our results showed that all the diagnostic kits evaluated were able to detect HBsAg irrespective of HBV genotypes. At the same time, it is apparent that some, but not all of the kits showed clear differences in sensitivity to the three HBV genotypes.     

Monoclonal Antibodies to Hepatitis B Surface Antigen (HBsAg) as a Tool for the Automated HBsAg Screening

Abstract. Hybridomas secreting monoclonal antibodies to hepatitis B surface antigen (HBsAg) were established by fusion of the spleen cells from mice immunized with purified HBsAg with the mouse myeloma cell line P3-NSI/l-Ag4-l. The monoclonal antibodies to the group-specific antigen (a) produced by one of them were used for the automated screening of HBsAg on the Groupamatic 360 (Kontron International). Its sensitivity is almost equal, but slightly inferior, to the system employing polyclonal horse antibodies to HBsAg; it barely detects 6 ng/ml of HBsAg. It is also as highly specific as the system with polyclonal antibodies; the incidence of false-positive reactions is 0.2%. These results indicate that the monoclonal antibodies will become a practical source for the HBsAg screening on the Groupamatic.     

Evaluation of rapid diagnostic tests for the detection of human immunodeficiency virus types 1 and 2, hepatitis B surface antigen, and syphilis in Ho Chi Minh City, Vietnam

An evaluation of three new rapid diagnostic test kits for human immunodeficiency virus types 1 and 2 (HIV-1/2), hepatitis B surface antigen (HBsAg), and syphilis involved a two-phase comparison of rapid diagnostic assays using prospectively collected from hospitals and clinics in Ho Chi Minh City, Vietnam. After specificity and sensitivity testing, three new rapid diagnostic test kits were tested in parallel with six commonly used diagnostic test kits. The Determine HIV-1/2 test had fewer indeterminate or equivocal results than the Capillus HIV-1/HIV-2 or HIV Blot 2.2 tests. However, the Determine HIV-1/2 test yielded one false-positive result when compared with the Serodia HIV, HIV Blot 2.2, and microparticle enzyme immunoassay (IMx) HIV tests. The Serodia HBsAg test yielded more false-negative results when compared with the Determine HBsAg diagnostic test kit. The results of the syphilis diagnostic tests evaluated in this clinical trial consistently agreed with those of the rapid plasma reagin test for syphilis. The Determine Syphilis Treponema pallidum (TP) test had three false-positive results compared with the Serodia TP and the Serodia TP x particle agglutination (PA) tests, which had two false-positive results that were confirmed as negative by an ELISA. Application of these serologic tests within this comparative evaluation framework, using the World Health Organization alternative testing strategies, proved to be an effective way to determine serostatus related to HIV, hepatitis B, and syphilis.     

Early detection of hepatitis B surface antigen and detection of HBsAg mutants: a comparison of five assays

BACKGROUND AND OBJECTIVES: Assays for the detection of hepatitis B surface antigen (HBsAg) face two challenges: the emergence of sensitive techniques for detection of hepatitis B virus DNA and the existence of HBsAg mutant hepatitis B viruses. We studied the sensitivity of five modern assays for detection of HBsAg. MATERIALS AND METHODS: The sensitivity of two mini robot-based assays--IMx HBsAg V2 and Vidas HBsAg--and three enzyme-linked immunosorbent assays (ELISAs) with microtitre plate format--Hepanostika Uni-Form II v1.2, Monolisa Ag HBs Plus and HBsAg Test System 3--was compared testing 11 HBsAg seroconversion series, serial dilutions prepared from three HBsAg standards and nine HBsAg mutant samples. RESULTS AND CONCLUSION: Overall, the IMx HBsAg V2 assay showed the highest sensitivity. It outperformed the Vidas HBsAg in analytical sensitivity and in detection of HBsAg mutants. Among the microtitre plate ELISAs, the Monolisa Ag HBs Plus outperformed the HBsAg Test System 3 in detection of HBsAg mutants and it surpassed the Hepanostika Uni-Form II v1.2 in analytical sensitivity.     

Specificities of serum alpha-fetoprotein in HBsAg+ and HBsAg- patients in the diagnosis of hepatocellular carcinoma.

Serum alpha-fetoprotein level is often elevated in patients with chronic liver disease and patients with hepatocellular carcinoma. One of the most difficult problems frequently encountered in practice is differentiating hepatocellular carcinoma from chronic liver disease. This study investigated the specificity and predictive value positive of serum alpha-fetoprotein at various levels in the diagnosis of hepatocellular carcinoma, using 54 patients with histologically proven hepatocellular carcinoma and 200 patients with chronic liver disease (40 patients with chronic active hepatitis and 160 patients with cirrhosis) as nontumor controls. Among 254 patients, 170 (66.9%) were HBsAg+. A wide range of overlap (from 0 to 6,400 ng/ml) in the distribution of serum alpha-fetoprotein levels between hepatocellular carcinoma and chronic liver disease patients was observed mainly among HBsAg+ patients. In contrast, the overlapping range of serum alpha-fetoprotein levels between HBsAg- patients with hepatocellular carcinoma and chronic liver disease was remarkably narrow (from 0 to 200 ng/ml). Therefore the specificity and predictive value positive of alpha-fetoprotein at a given level were significantly lower in HBsAg+ than in HBsAg- patients, especially when alpha-fetoprotein was between 25 and 200 ng/ml. The specificities of alpha-fetoprotein at 200 ng/ml and 400 ng/ml in HBsAg+ patients were 79.8% and 91.5%, respectively, whereas these specificities were both 100% in HBsAg- patients. The predictive values positive at 200 ng/ml and 400 ng/ml in HBsAg+ patients were 53.6% and 72.5%, respectively, in contrast to 100% at both levels in HBsAg- patients. The serum alpha-fetoprotein level, which showed a predictive value positive of 95% in HBsAg+ hepatocellular carcinoma patients, was 3,200 ng/ml, whereas that in HBsAg- hepatocellular carcinoma patients, was 200 ng/ml. We conclude that serum HBsAg status should be considered when serum alpha-fetoprotein is measured as an independent test to diagnose hepatocellular carcinoma, and suggest that regular serum alpha-fetoprotein determination may be more useful in HBsAg- patients with chronic liver disease for the early diagnosis of hepatocellular carcinoma than in HBsAg+ patients.     

乙型肝炎表面抗原定性测定室内质控物浓度的选择方法

目的 建立定性酶联免疫吸附法(ELISA)测定室内质量控制质控物浓度选择和Levey-Jennings质控图在不同批试剂间连续作图的方法。 方法 以乙型肝炎表面抗原(HBsAg)的ELISA测定为例,先按NCCLs EP5文件评价方法对系列质控血清(含量分别为0.2、0.5、1.0、2.0和5.0 ng/ml)的批内和批间测定精密度,并根据任一次测定值(S/CO)最好的线性相关确定回归直线方程(y=bx a),选择计算得到的S/CO值减去精密度测定中得到的3倍批间CV％与该S/CO值的乘积(意即3倍SD)仍大于1的质控物,作为室内质控使用。不同批试剂间质控图的连续,则在换用新批号试剂时,用新批号试剂测定上述系列质控血清,得到一新的直线方程(y2=b2x2 a2)。原批号试剂同样也可得一直线方程(y1=b1x1 a1)。根据两个直线方程即可得到y2/y1之间的换算因子,从而可以将新批号试剂对相同室内质控血清的测定值换算回去,在原质控图上继续作图。结果 所用的HBsAg ELISA测定方法测定含量分别为0.2、0.5、1.0、2.0和5.0ng/ml质控血清的批内变异分别为11.08％、9.49％、9.83％、9.18％和7.25％,批间变异分别为13.25％、14.03％、15.11％、13.29％和9.92％。任选一次测定的具有最好相关的回归直线方程y=3.509x 0.180,可选择的室内质控血清浓度可为0.5或1.0ng/ml。换用其它两批     

Binding activity of HBsAg particles from chronic HBsAg carriers to polystyrene beads coated with polymerized human serum albumin: diagnostic relevance of the assay

The binding activity of HBsAg particles to polystyrene beads coated with polymerized human serum albumin (pHSA) was studied by radioimmunoassay in 48 patients with chronic HBsAg carrier state. The pHSA assay was positive in all 16 HBeAg-positive patients and in 22 HBeAg-negative HBsAg carriers with chronic hepatitis. Asymptomatic, "healthy" HBsAg carriers did not react in the pHSA assay. Mean binding activity was significantly higher in the HBeAg-positive group (P/N ratio 39.3) than in HBeAg-negative carriers with chronic hepatitis in various stages (P/N ratio 19.2). Fractionation of five representative HBeAg-positive sera by density gradient ultracentrifugation in cesium chloride yielded three peaks of HBsAg particles at 1.28, 1.22 and. 1.18 g/ml. The first HBsAg peak contained Dane particles and exhibited strong reactivity in pHSA assay. The second and third peaks, both consisting of 22 nm particles, reacted differently in pHSA assay. While about half of the HBsAg particles in the second peak were bound, reaction in the third HBsAg peak was predominantly negative. Intrahepatic HBsAg was detectable with the immunofluorescence technique in 19 of 22 HBsAg carriers. HBcAg was found in seven of 10 HBeAg-positive cases and in three of 16 HBeAg-negative patients with chronic hepatitis. The diagnostic value of pHSA assay might be seen in differentiation between HBeAg-negative chronic HBsAg carriers with liver disease or "healthy" carrier state.     

Molecular characterization of hepatitis B virus surface antigen mutants in Singapore patients with hepatocellular carcinoma and hepatitis B virus carriers negative for HBsAg but positive for anti‐HBs and anti‐HBc

Background and Aims : Mutations on the a‐determinant of hepatitis B virus surface antigen (HBsAg), capable of escaping detection and vaccination, are identified in HBsAg‐positive/anti‐HBs‐positive vaccinated infants. We studied the prevalence of these mutants in HBsAg‐negative/anti‐HBc‐positive chronic HBV carriers and patients with hepatocellular carcinoma (HCC). Methods : DNA sequence coding for the antigenic a‐determinant of HBsAg was amplified from either HCC genomic DNA or serum samples of the selected patients and sequenced. The replicative mutant genomes were reconstituted in vitro and their reactivity to commercial kits measured. Results : Mutations within and/or outside the a‐determinant were identified in patients seronegative for HBsAg. They were then reconstituted in vitro and transiently transfected into HepG2 cells. Culture medium containing secreted HBV viral particles was collected and assayed for their binding to commercial kits. Drastic decrease of reactivity to these kits was seen with most of the identified mutations, including those located outside the a‐determinant. Conclusion : The existence of a more complex antigenic structure of HBsAg is indicated by the decreased reactivity to detection of mutations, some of which are outside the a‐determinant, escape vaccination and may persist in seronegative patients. The high proportion of HBsAg mutants that are integrated in HCC genomes suggests a role of these mutants in hepatocarcinogenesis, possibly leading to mutant HBV‐related HCC. © 2002 Blackwell Publishing Asia Pty Ltd     

Relevance of antibody content and test format in HIV testing of pooled sera.

OBJECTIVE: To determine the effects of HIV-1 antibody level and test-format characteristics on testing pooled sera. DESIGN: This study was designed with a laboratory exercise followed by test observations on serosurveillance samples. METHODS: Sera with low, medium and high (n = 22, 12 and 20, respectively) antibody titers were pooled with HIV-1-negative sera and tested with two enzyme-linked immunosorbent assays (ELISA) and a particle agglutination test. The same kits were used to test single and pooled (batches of five, 10 and 20) samples collected from 3000 blood donors and sex workers. These samples were then seeded with 50 varying antibody-containing sera and similarly tested. Initial reactivities, sensitivities, and specificities for all test kits were calculated and compared. RESULTS: In the laboratory exercise, all reactive pools of five were detected. False-negative pools in batches of 10 and 20 with low antibody titers were noted with one or both ELISA, but not with the particle agglutination method. Testing 3000 samples revealed three confirmed reactive samples and 100% sensitivity/specificity for all kits, for both single and pooled sera testing. Increased initial reactivity (IR) was noted for the two ELISA. Examinations of pools of the seeded 3000 samples with the two ELISA showed false-negative reactivity with pools of 10 and 20 when pools contained low antibody sera (sensitivities and specificities of 92-97.9% and 98.1-100%, respectively). Again, increased IR was seen with the ELISA. False-negative pool and increased IR was not seen with the agglutination test (sensitivity/specificity 100%). CONCLUSIONS: We recommend the use of the particle agglutination assay for testing pooled sera of batches of 20 or less. Components of reactive pools should then be tested and reactive samples should undergo supplementary testing. Pooled samples tested by ELISA should not exceed five per batch. Retesting of reactive pools, testing of its components, and supplemental test(s) of reactive sera should then follow. The optimum pool size for most laboratories is five, with the best technical and economic performance seen with the particle agglutination assay.     

Serum alpha-fetoprotein for diagnosis of hepatocellular carcinoma in patients with chronic liver disease: influence of HBsAg and anti-HCV status

BACKGROUND: It is not established whether virological status affects the efficiency of alpha-fetoprotein (AFP) as a hepatocellular carcinoma (HCC) marker among patients with chronic liver disease (CLD). METHODS: We enrolled in a case-control study 170 HCC and 170 CLD patients, matched for age, sex, CLD and HBsAg/anti-HCV status. The AFP sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were calculated. PPV and NPV were evaluated for three additional HCC prevalences (5, 10, and 20%). RESULTS: The best discriminating AFP value was 16 ng/ml. A value of 20 ng/ml (above which investigations for HCC are recommended) had equivalent sensitivity (60.0 vs. 62.4%) and specificity (90.6 vs. 89.4%). PPV of 20 ng/ml was 84.6% but decreased to 25.1% at 5% tumor prevalence. NPV was 69.4% and rose to 97.7% at 5% prevalence. In the different groups of infected patients PPV ranged from 80.0 to 90.9%, falling to 17.4-34.5% at 5% prevalence. In noninfected patients PPV was 100% at any HCC prevalence. NPV ranged from 59.0 to 73.0%, reaching 96.5-98.1% at 5% prevalence. CONCLUSIONS: In CLD patients, AFP monitoring misses many HCCs and inappropriately arouses suspicion of malignancy in many patients. Its usefulness is barely affected by the infection responsible for CLD. An AFP elevation could be more indicative of HCC in non-infected patients.     

Clinical features of hepatocellular carcinoma seronegative for both HBsAg and anti-HCV antibody but positive for anti-HBc antibody in Japan

OBJECTIVE: We determined the prevalence of patients with hepatocellular carcinoma (HCC) who were positive for only anti-hepatitis B core (anti-HBc) antibody among 284 Japanese patients and compared their clinical features to those who were hepatitis B surface antigen positive [HBsAg(+)]. METHODS: Serum HBsAg and anti-hepatitis C virus (anti-HCV) antibody were examined for all HCC patients. Testing for anti-HBc antibody was performed in the HBsAg(-)/anti-HCV(-) patients. The clinical factors and the survival rate were compared between the HBsAg(+) patients (HCC-B) and those positive for anti-HBc alone (HCC-PB). RESULTS: There were 19 (6.7%) HBsAg(+), 236 (83.1%) anti-HCV(+), seven (2.5%) HBsAg(+)/anti-HCV(+), and 22 (7.7%) HBsAg(-)/anti-HCV(-) among the 284 patients tested. Sixteen (72.7%) of the 22 HBsAg(-)/anti-HCV(-) patients were assigned to the HCC-PB group. The prevalence of positivity for anti-HBc alone among all 284 HCC patients was 5.6%. Significant differences between the HCC-PB and HCC-B groups were that age at diagnosis was higher in the HCC-PB group (72.1 yr) than in the HCC-B group (56.2 yr) (p < 0.001), the serum alpha-fetoprotein concentrations were lower in the HCC-PB group (8.2 ng/ml) than in the HCC-B group (43 ng/ml) (p = 0.0488), and a higher familial history of liver disease was observed in the HCC-B group (p = 0.0373). However, there was no significant difference in the cumulative survival rate. CONCLUSIONS: Positivity for anti-HBc alone is not rare compared to HBsAg(+), and the clinical features of positivity for anti-HBc alone are similar to those of HBsAg(+). Some differences in the clinical features between the two groups may be explained by differences in the time of first exposure to hepatitis B virus. Therefore, the natural course of acute hepatitis B may be reconsidered.     

五种布鲁氏菌核酸检测试剂盒检测性能的评价北大核心CSCD

目的比较5种布鲁氏菌核酸实时荧光PCR检测试剂盒的一致性和检出能力,为临床实验室选择检测方法和布鲁氏菌的诊断提供参考依据。方法选用经病原学检测确定为布鲁氏菌阳性的血液样本38份,健康人的血液样本24份,潘氏变形杆菌、溶藻弧菌、河弧菌、铜绿假单胞菌、肺炎克雷伯菌DNA各1份,使用5种试剂盒(编号A-E)分别进行核酸检测,比较5种试剂盒临床样本检测的一致性;选择1份阳性样本核酸用无RNA酶水梯度稀释得到5个浓度(浓度1:4453.13 fg/μL,浓度2:1113.28 fg/μL,浓度3:278.32 fg/μL,浓度4:69.58 fg/μL,浓度5:17.40 fg/μL),每个浓度使用5种试剂盒(编号A-E)分别进行3次检测,比较5种试剂盒的阳性检出率及批内重复性。结果5种试剂盒检测67份DNA样品的符合率稍有不同,试剂盒ABDE的符合率均为100%,试剂盒C的符合率为98.51%。批内重复性显示5种试剂盒在浓度1、浓度2、浓度3水平重复检测DNA的Ct值变异系数均<5%;在浓度1与浓度4梯度区间,试剂盒的阳性检出能力比较显示试剂盒A、B、D较高,为11/12,试剂盒C和E较低,为8/12。结论5种试剂盒的真实性和可靠性较好,灵敏度和符合率稍有差别,特异度均为100%;重复性较好,检测性能良好。部分试剂盒对弱阳性样本的检出能力不强,该类样本可使用多种试剂盒复核,以保障结果的准确性。     

血清HBsAg阳性孕妇血清细胞因子水平对HBV母婴传播的影响

目的 探讨血清干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)和IL-10水平对乙型肝炎病毒(HBV)母婴传播的影响。方法 2017年8月～2019年7月我院诊治的126例血清HBsAg阳性孕妇和55例健康孕妇,采用ELISA法检测孕妇和新生儿血清IFN-γ、IL-4、IL-10水平。采用受试者工作特征(ROC)曲线和Logistic回归模型分析孕妇外周静脉血细胞因子水平对HBV母婴传播的影响。结果 本组发生HBV宫内感染15例,自宫内未感染者中选择30例作为对照,结果宫内感染组孕妇外周静脉血IFN-γ水平为(681.3±141.6)pg/ml,IL-10水平为(62.3±11.4)pg/ml,显著低于宫内未感染组【分别为(992.6±192.3)pg/ml和(68.5±12.8)pg/ml】或对照组【分别为(1040.4±48.1)pg/ml和(73.7±2.6)pg/ml,P＜0.05】,而血清IL-4水平为(68.1±22.9)ng/L,显著高于宫内感染组或对照组【分别为(49.2±15.3)ng/L和(38.9±7.2)ng/L,P＜0.05】;三组新生儿脐静脉血三个细胞因子水平无显著性差异(P＞0.05);ROC分析显示,孕妇外周静脉血IFN-γ预测HBV母婴传播的曲线下面积(AUC)为0.877(95%CI:0.810～0.945),诊断的截断点为≤782.36 pg/ml,其敏感度为100.0%,特异度为69.4%。结论 HBV感染孕妇存在免疫功能调节紊乱,检测血清IFNγ、IL-4和IL-10等细胞因子水平可作为评估HBV母婴传播的观察指标,能为临床确定防治措施提供参考信息。     

Detection of hepatitis B virus markers using a biosensor based on imaging ellipsometry

Summary.  A biosensor based on imaging ellipsometry (BIE) has been developed and validated in 169 patients for detecting five markers of hepatitis B virus (HBV) infection. The methodology has been established to pave the way for clinical diagnosis, including ligand screening, determination of the sensitivity, set-up of cut-off values (CoVs) and comparison with other clinical methods. A matrix assay method was established for ligand screening. The CoVs of HBV markers were derived with the help of receiver operating characteristic curves. Enzyme-linked immunosorbent assay (ELISA) was the reference method. Ligands with high bioactivity were selected and sensitivities of 1 ng/mL and 1 IU/mL for hepatitis B surface antigen (HBsAg) and surface antibody (anti-HBs) were obtained respectively. The CoVs of HBsAg, anti-HBs, hepatitis B e antigen, hepatitis B e antibody and core antibody were as follows: 15%, 18%, 15%, 20% and 15%, respectively, which were the percentages over the values of corresponding ligand controls. BIE can simultaneously detect up to five markers within 1 h with results in acceptable agreement with ELISA, and thus shows a potential for diagnosing hepatitis B with high throughput.     

Serum hepatitis B virus DNA levels and liver histology in inactive HBsAg carriers

BACKGROUND/AIMS: A recent NIH research workshop on hepatitis B virus (HBV) revisited the definition of healthy HBsAg carriers. The new definition inactive surface antigen (HBsAg) carriers includes an estimated serum HBV DNA level below 105 copies/ml. However, this cut-off value needs to be confirmed. METHODS: Eighty-five consecutive patients, HBsAg-positive/HBeAg-negative with persistently normal alanine aminotransferase (ALT) and undetectable serum HBV DNA with standard assay (Versant HBV DNA Assay (bDNA), Bayer) were prospectively followed for 3.2+/-2.6 (range 0.5-11) years; 58 underwent a liver biopsy. Serum HBV DNA was quantified with a sensitive polymerase chain reaction assay (Cobas Amplicor HBV Monitor, Roche) (sensitivity 200 copies/ml), and liver histology was assessed using the Ishak scoring system. RESULTS: The median serum HBV DNA level was 1300 copies/ml (<200-179 x 10(3) copies/ml), 16% of the subjects had no detectable serum HBV DNA and 98% had levels below 10(5) copies/ml. Histologic lesions were mild (total score <7) in all cases. Loss of HBsAg was observed in three patients, three patients experienced a transient increase in ALT (<2 x upper limit of normal), and serum HBV DNA levels remained stable (1-6 years) in 97% of the 38 patients retested. CONCLUSIONS: In our study of inactive HBsAg carriers, the median serum HBV DNA level was 1300 copies/ml, the serum HBV DNA level was below 10(5) copies/ml in 98% of the patients, and remained stable; histological lesions were mild in all cases.     

Diagnostic value of serum ferritin in primary hepatocellular carcinoma

The diagnostic value of serum ferritin levels was evaluated in 19 patients with biopsy-proven primary hepatocellular carcinoma (PHC) and 26 patients with chronic liver disease (CLD). Serum ferritin levels were significantly elevated in PHC, as compared with CLD and controls (p less than 0.0005). Similarly, serum ferritin/SGOT ratio, an index of increased ferritin production, was significantly higher in PHC than in CLD and controls. Serum alpha-fetoprotein (alpha-FP) was higher in PHC than in CLD (p less than 0.0025). No significant correlation was noted between serum ferritin and alpha-fetoprotein or SGOT in PHC and CLD. 17 of 19 patients with PHC had serum ferritin values over 450 ng/ml (sensitivity 88%). By contrast, only 10 of 17 patients with PHC (59%) demonstrated alpha-FP levels over 25 ng/ml, compatible with the diagnosis of PHC. 9 of these 10 patients had ferritin levels over 450 ng/ml, within the distribution of values for PHC. Conversely, 7 of 17 patients with PHC (40%) had normal levels of alpha-FP (false-negative). However, 6 of these patients (86%) had ferritin levels over 450 ng/ml, consistent with values in PHC. In this study, the overall sensitivity of serum ferritin in PHC was higher than that of alpha-FP (88 versus 59%) and its specificity 85 versus 68% for alpha-FP. These data indicate that serum ferritin may be utilized as a useful diagnostic marker in the evaluation of patients with PHC.     

HBsAg level at time of liver transplantation determines HBsAg decrease and anti-HBs increase and affects HBV DNA decrease during early immunoglobulin administration

BACKGROUND/AIMS: Administration of hepatitis B immunoglobulin (HBIG) initially after liver transplantation of hepatitis B patients is considered important to prevent reinfection reliably. However, dosing schedules differ considerably between centers. We measured HBsAg, anti-HBs and HBV DNA kinetics to create a rational basis for dosing schemes. METHODS: Thirteen patients (group A) received 10,000 IU HBIG in the anhepatic phase followed by 10,000 IU daily until HBsAg became negative, whereas five patients (group B) received 20,000 IU followed by 5000 IU every 30 min. RESULTS: HBsAg levels at time of transplantation ranged from 0.12 to 12,990 IU/ml. Correlations between initial HBsAg and HBIG required to decrease HBsAg below 1 IU/ml were high in groups A and B (r=0.97, p<0.001; r=1.00, p<0.001), as were correlations between initial HBsAg and HBIG required to raise anti-HBs above 1000 IU/l (r=0.94, p<0.001; r=1.00, p<0.001). In 11 HBV DNA-positive patients, DNA levels became negative in seven, and dropped by 2.5 log10 (mean) in the other four patients during immunoglobulin administration. CONCLUSIONS: In conclusion, required HBIG doses to decrease HBsAg and raise anti-HBs are determined by HBsAg levels at time of transplantation, not by HBV DNA levels. Shortened HBIG dosing intervals accelerate HBsAg decrease and anti-HBs increase. HBV DNA decreases rapidly during HBIG administration in most patients.     

Detection, by three techniques, of hepatitis B surface antigen (HBsAg) and determination of HBsAg and anti-HBs titres in patients with chronic liver disease.

The sensitivities of three technqiues used to detect serum hepatitis B surface antigen (HBsAg) were compared in 411 patients with various types of chronic liver disease. Counterimmunoelectrophoresis proved an unreliable test. Two haemagglutination technqiues were slightly less sensitive than radioimmunoassay but were more rapidly performed. Less sensitive techniques were particularly unreliable in active liver disease where HBsAg titres were low. HBsAg was detected in patients with chronic persistent hepatitis, alcoholic liver disease, chronic active liver disease with or without cirrhosis, and primary liver cell carcinoma. Forty-six of the 68 (68%) HBsAg positive subjects were males coming from outside the United Kingdom. The HBsAg titres in 13 subjects with chronic persistent hepatitis were significantly higher (P less than 0-001) than those in 43 subjects with chronic active liver disease. Corticosteroid therapy did not alter the HBsAg titre significantly. None of the 28 HBsAg positive subjects studied serially for up to two years cleared HBsAg from the serum. Anti-HBs was examined by passive haemagglutination and found in 35 subjects, 26 of whom had no evidence of liver disease, 80% came from abroad. Anti-HBs was believed to be of epidemiological rather than of pathological importance.     

Midnight salivary cortisol for the initial diagnosis of Cushing's syndrome of various causes

We assessed the value of midnight salivary cortisol for the initial diagnosis of Cushing's syndrome. Sixty-three patients with various causes of Cushing's syndrome (37 with Cushing's disease, 17 with adrenal Cushing's syndrome, and nine with ectopic ACTH syndrome) and 54 control subjects with simple obesity were studied. All patients with Cushing's syndrome excreted more than 90 microg urinary free cortisol (UFC)/d (248 nmol/d), and all controls excreted less than 90 microg/d UFC. All patients with Cushing's syndrome had a midnight salivary cortisol concentration above 2.0 ng/ml (5.52 nmol/liter), whereas only three controls did so [2.0 ng/ml (5.52 nmol/liter); 2.05 ng/ml (5.66 nmol/liter); and 3.6 ng/ml (9.96 nmol/liter)]. This cut-off provides a sensitivity of 100% and a specificity of 96%. In patients with Cushing's syndrome, midnight salivary cortisol concentrations were correlated with UFC collected over the same period of time (0800-0800 h). Salivary cortisol measurements taken every 4 h showed a typical lack of circadian variation. The daily measurement of midnight salivary cortisol concentrations for 2 wk or more in five other out-patients (with obvious Cushing's disease, subclinical adrenal Cushing's syndrome, suspected Cushing's syndrome, pituitary incidentaloma, and prolactinoma) demonstrated the clinical utility of this factor. Measuring midnight salivary cortisol is an easy and noninvasive means of diagnosing hypercortisolism. Its diagnostic accuracy is identical to, if not better than, that of previously described gold standards.     

sICAM-1 as a serum marker in the diagnosis and follow-up of treatment of pulmonary tuberculosis.

OBJECTIVE: To detect the value of sICAM-1 in the diagnosis and follow-up of treatment of tuberculosis. DESIGN: sICAM-1 levels were evaluated before and after treatment in 30 patients with pulmonary tuberculosis, only before treatment in five patients with pneumonia, five with lung cancer, and five with bronchiectasis, and in 10 healthy volunteers. RESULTS : sICAM-1 levels were as follows: 436.2 +/- 194.4 ng/ml in patients with pulmonary tuberculosis, 274 +/- 32.1 ng/ml in lung cancer patients, 268 +/- 41.9 ng/ml in bronchiectasis patients, 199.6 +/- 43.1 ng/ml in pneumonia patients, and 146.5 +/- 20.2 ng/ml in healthy individuals. sICAM-1 levels of tuberculosis cases before treatment were higher than in both the healthy group and in all the other groups. The levels in the healthy group were lower than in all other groups. CONCLUSION: The cut-off point (298 ng/ml) obtained by adding the standard deviation to the mean sICAM-1 value of patients without tuberculosis had 83.3% sensitivity, 86.6% specificity and 84.4% accuracy in differentiating pulmonary tuberculosis from other pulmonary diseases. sICAM-1 can be used as an auxiliary marker in the diagnosis of pulmonary tuberculosis.